A single-tube duplex and multiplex PCR for simultaneous detection of four cassava mosaic begomovirus species in cassava plants

► We developed a single-tube PCR for simultaneous detection of cassava begomoviruses. ► Developed assay detected four CMBs in uniplex, duplex and multiplex PCR. ► Protocol modified to yield high quality DNA from dry-stored cassava leaf samples. ► The multiplex PCR can be used as a rapid test for cassava mosaic begomoviruses.